Disposable sensors for rapid screening of mutated genes

Analytical and Bioanalytical Chemistry
T GarcíaE Lorenzo

Abstract

A screening method for rapid detection of gene mutations directly in polymerase chain reaction (PCR) of genomic DNA is described. The method involves the development of a disposable screen-printed gold electrode modified with a thiolated capture probe directly obtained from denaturated PCR genomic DNA, which recognizes (by hybridization) its fully complementary sequence (wild type), giving a signal, whereas no signal is obtained for single-mismatched target (mutant). The detection of the hybridization event is achieved by changes in the metal redox center electroactivity of the complex [Ru(NH(3))(5) L](2+), where L is [3-(2-phenanthren-9-yl-vinyl)-pyridine], at -0.200 V. This complex binds to double-stranded DNA in a very selective form. The method allows discrimination between the wild type and the mutant of gene MRP3 directly in large PCR amplicons extracted from blood cells, without the need to use either synthetic probes or labeled targets. The mutation involves the presence of a single-nucleotide polymorphism (SNP) at base 54 of a 145-base-pair sequence from exon 21 of gene MRP3. Since the presence of this SNP might lead to a variety of hereditary liver disorders, its identification in a rapid and easy form may provide nov...Continue Reading

References

Oct 12, 2000·Analytical Chemistry·D W Pang, H D Abruña
Feb 26, 2003·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Ramón BatallerDavid A Brenner
Dec 20, 2003·Biosensors & Bioelectronics·Fausto LucarelliMarco Mascini
Dec 30, 2004·Analytical Chemistry·Natalia C TansilZhiqiang Gao
Jan 1, 2005·Biosensors & Bioelectronics·M V Del PozoE Lorenzo
Jun 3, 2005·Nature Reviews. Drug Discovery·Neil Kaplowitz
Jun 25, 2005·Journal of Hepatology·Christiane Pauli-MagnusPeter J Meier
Feb 24, 2006·Alcohol and Alcoholism : International Journal of the Medical Council on Alcoholism·Felix Stickel, Christoph H Osterreicher

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Citations

Mar 21, 2016·Applied Microbiology and Biotechnology·Caspar DemuthDieter Eibl
Sep 13, 2020·Biosensors·Emiliano Martínez-PeriñánEncarnación Lorenzo
Oct 31, 2021·Mikrochimica Acta·Laura Gutiérrez-GálvezEncarnación Lorenzo

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